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Journal of Dental Research
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Biological

Mouse Amelogenin Exons 8 and 9: Sequence Analysis and Protein Distribution

P. Papagerakis, J.M. Ibarra, N. Inozentseva, P. DenBesten1 and M. MacDougall*

Department of Pediatric Dentistry, Dental School, University of Texas Health Science Center San Antonio, MSC 7888, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900, USA; and
1 Department of Growth & Development, Pediatric Dentistry Division, School of Dentistry, University of California San Francisco, San Francisco, CA;

Correspondence: * corresponding author, macdougall{at}uthscsa.edu

Amelogenin is the major protein of the developing enamel. Two additional exons, termed 8 and 9, have been characterized in the rat. Our aim was: to identify the mouse amelogenin exons 8/9 sequences; to investigate the potential presence of the alternative spliced isoforms of amelogenin exons 8/9; and to immunolocalize proteins containing sequences encoded by exons 8/9 during odontogenesis. RT-PCR analysis with exon 9 anti-sense primer generated 2 major amplicons with the use of a mouse tooth cDNA library and dental cell lines. DNA sequence analysis showed 93% identify with the rat exons 8/9 sequence. Alternative splicing of exon 3 was also found, but only in cDNAs lacking exons 8 and 9. Immunohistochemistry localized exons 8/9-encoded proteins in ameloblasts, young odontoblasts, and stratum intermedium cells. Analysis of our data supports the hypothesis that: (1) AMELX contains 2 additional exons; (2) ameloblasts and odontoblasts synthesize amelogenin 8/9; and (3) amelogenin splice variants may have unique functions during tooth formation.

Key Words: amelogenin • ameloblasts • odontoblasts • alternative splicing • exons 8 and 9

Journal of Dental Research, Vol. 84, No. 7, 613-617 (2005)
DOI: 10.1177/154405910508400706
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